Cover Caption: The brittle Mutant of Rice
With the rigid support from the cell walls, plants are
able to stand up and grow upwards. The recessive
brittle mutant of rice (Oryza sat iva L. subsp. indica)
was obtained after EMS-mutagenesis from the cultivar
of E-you 532. The mutation affected the cell wall
biosynthesis and caused the reduction of cell length
and cell wall thickness, which led to an alteration of
other phenotypes as well. The BRITTLE gene is
located on the chromosome 10. See pages 319每328
for details.

Bioenergy Plants

Water Sources of Dominant Species in Three Alpine Ecosystems on the Tibetan Plateau, ChinaAuthor: De-Yu Duan, Hua Ouyang, Ming-Hua Song and Qi-Wu Hu
Journal of Integrative Plant Biology 2008 50(3): 257-264DOI: 10.1111/j.1744-7909.2007.00633.x

Plant water sources were estimated by two or three compartment linear mixing models using hydrogen and oxygen isotope (汛D and 汛18O) values of different components such as plant xylem water, precipitation and river water as well as soil water on the Tibetan Plateau in the summer of 2005. Four dominant species (Quercus aquifolioides, Pinus tabulaeformis, Salix rehderiana and Nitraria tangutorum) in three typical ecosystems (forest, shrub and desert) were investigated in this study. Stable isotope ratios of the summer precipitations and the soil water presented variations in spatial and temporal scales. 汛18O values of N. tangutorum xylem water were constant in the whole growth season and very similar to those of deep soil water. Water sources for all of the plants came from both precipitations and soil water. Plants switched rapidly among different water sources when environmental water conditions changed. Rainwater had different contributions to the plants, which was influenced by amounts of precipitation. The percentage of plant xylem water derived from rainwater rose with an increase in precipitation. Water sources for broad-leaved and coniferous species were different although they grew in the same environmental conditions. For example, the broad-leaved species Q. aquifolioides used mainly the water from deep soil, while 92.5% of xylem water of the coniferous species P. tabulaeformis was derived from rainwater during the growth season. The study will be helpful for us to fully understand responses of species on the Tibetan Plateau to changes in precipitation patterns, and to assess accurately changes of vegetation distribution in the future.

Growth and wood and bark properties of Abies faxoniana seedlings after one year's exposure to elevated CO2 concentration (ambient + 350 (㊣ 25) 米mol/mol) under two planting densities (28 or 84 plants/m2) were investigated in closed-top chambers. Tree height, stem diameter and cross-sectional area, and total biomass were enhanced under elevated CO2 concentration, and reduced under high planting density. Most traits of stem bark were improved under elevated CO2 concentration and reduced under high planting density. Stem wood production was significantly increased in volume under elevated CO2 concentration under both densities, and the stem wood density decreased under elevated CO2 concentration and increased under high planting density. These results suggest that the response of stem wood and bark to elevated CO2 concentration is density dependent. This may be of great importance in a future CO2 enriched world in natural forests where plant density varies considerably. The results also show that the bark/wood ratio in diameter, stem cross-sectional area and dry weight are not proportionally affected by elevated CO2 concentration under the two contrasting planting densities. This indicates that the response magnitude of stem bark and stem wood to elevated CO2 concentration are different but their response directions are the same.

From 30 June to 24 September in 2003 ecosystem respiration (Re) in two alpine meadows on the Tibetan Plateau were measured using static chamber- and gas chromatography- (GC) based techniques. Simultaneously, plant removal treatments were set to partition Re into plant autotrophic respiration (Ra) and microbial heterotrophic respiration (Rh). Results indicated that Re had clear diurnal and seasonal variation patterns in both of the meadows. The seasonal variability of Re at both meadow sites was caused mainly by changes in Ra, rather than Rh. Moreover, at the Kobresia humilis meadow site (K_site), Ra and Rh accounted for 54% and 46% of Re, respectively. While at the Potentilla fruticosa scrub meadow (P_site), the counterparts accounted for 61% and 39%, respectively. T test showed that there was significant difference in Re rates between the two meadows (t = 2.387, P = 0.022). However, no significant difference was found in Rh rates, whereas a significant difference was observed in Ra rates between the two meadows. Thus, the difference in Re rate between the two meadows was mainly attributed to plant autotrophic respirations. During the growing season, the two meadows showed relatively low Q10 values, suggesting that Re, especially Rh was not sensitive to temperature variation in the growing season. Additionally, Re and Rh at the K_site, as well as Rh at the P_site was negatively correlated with soil moisture, indicating that soil moisture would also play an important role in respirations.

The self-thinning rule defines a straight upper boundary line on log-log scales for all possible combinations of mean individual biomass and density in plant populations. Recently, the traditional slope of the upper boundary line, −3/2, has been challenged by −4/3 which is deduced from some new mechanical theories, like the metabolic theory. More experimental or field studies should be carried out to identify the more accurate self-thinning exponent. But it's hard to obtain the accurate self-thinning exponent by fitting to data points directly because of the intrinsic problem of subjectivity in data selection. The virtual dynamic thinning line is derived from the competition-density (C-D) effect as the initial density tends to be positive infinity, avoiding the data selection process. The purpose of this study was to study the relationship between the virtual dynamic thinning line and the upper boundary line in simulated plant stands. Our research showed that the upper boundary line and the virtual dynamic thinning line were both straight lines on log-log scales. The slopes were almost the same value with only a very little difference of 0.059, and the intercept of the upper boundary line was a little larger than that of the virtual dynamic thinning line. As initial size and spatial distribution patterns became more uniform, the virtual dynamic thinning line was more similar to the upper boundary line. This implies that, given appropriate parameters, the virtual dynamic thinning line may be used as the upper boundary line in simulated plant stands.

The present study aims to identify the narrow spectral bands that are most suitable for characterizing rice biophysical parameters. The data used for this study come from ground-level hyperspectral reflectance measurements for five rice species at three levels of nitrogen fertilization during the growing period. Reflectance was measured in discrete narrow bands between 350 and 2 500 nm. Observed rice biophysical parameters included leaf area index (LAI), wet biomass and dry biomass. The stepwise regression method was applied to identify the optimal bands for rice biophysical parameter estimation. This research indicated that combinations of four narrow bands in stepwise regression models explained 69% to 83% variability for LAI, 56% to 73% for aboveground wet biomass and 70% to 83% for leaf wet biomass. An overwhelming proportion of rice information was in a particular portion of near infrared (NIR) (1 100每1 150 nm), red-edge (700每750 nm), and a longer portion of green (550每600 nm). These were followed by the moisture-sensitive NIR (950每1 000 nm), the intermediate portion of shortwave infrared (SWIR) (1 650每1 700 nm), and another portion of NIR (1 000每1 050 nm).

In order to analyse the adaptation potential of tomato shoots to a sudden increase in Cd concentration, tomato plants (Solanum lycopersicum L.; cultivar Ailsa Craig) were exposed under controlled environmental conditions to a high dose of this heavy metal (250 然 CdCl2) in nutrient solution for 7 and 14 days. Both root and shoot growth was completely inhibited but all plants remained alive until the end of the treatment. Cell viability remained unaffected but the activity of the unproductive mitochondrial alternative pathway was stimulated by Cd stress at the expense of the cytochrome pathway. Cadmium concentration was higher in roots than in shoots and a decrease in the rate of net Cd translocation was noticed during the second week of stress. Cadmium decreased both leaf conductance (gl) and chlorophyll concentration. Impact on net CO2 assimilation however remained limited and soluble sugars accumulated in leaves. Photochemical efficiency of PSII (Fv/Fm) was not affected despite a decrease in the number of reaction centers and an inhibition of electron transfer to acceptors of PSII. It is concluded that tomato shoot may sustain short term exposure to high doses of cadmium despite growth inhibition. This property implies several physiological strategies linked to both avoidance and tolerance mechanisms.

Twelve cassava landraces were evaluated for sources of resistance genes to diseases and pests of major economic importance in Africa. The objective was to assess their levels of field resistance to mosaic disease (ACMD), bacterial blight (CBB), anthracnose (CAD), and green mite (CGM), compared to TMS30572, an elite cultivar widely adopted in Africa. Considerable genotypic variation was observed among cultivars for resistance to ACMD and CGM but not for CBB and CAD. The lowest mean incidence of 12% and severity of 1.8 on a scale of 1-5 for ACMD was recorded for Atu, a landrace with farmer acceptable qualities. In comparison, the improved cultivar, TMS 30572, had a mean disease incidence of 72% and a severity score of 2.8. Another landrace, MS-20 had the lowest CGM damage score (2.1) while TMS 30572 emerged as one of the susceptible cultivar with a damage score of 3.0. Additional sources of resistance to ACMD and CGM that may possibly be better than the popular improved cultivar; TMS 30572 were identified in this study. These could serve as novel sources of additional genes to complement existing resources for elite cassava breeding in Africa.

Plant mechanical strength is an important agronomic trait of rice. An EMS-induced rice mutant, fragile plant 2 (fp2), showed morphological changes and reduced mechanical strength. Genetic analysis indicated that the brittle of fp2 was controlled by a recessive gene. The fp2 gene was mapped on chromosome 10. Anatomical analyses showed that the fp2 mutation caused the reduction of cell length and cell wall thickness, increasing of cell width, and the alteration of cell wall structure as well as the vessel elements. And the consequence was the global alteration in plant morphology. Chemical analyses indicated that the contents of cellulose and lignin decreased, hemicelluloses and silicon increased in fp2. These results were different from the other mutants reported in rice. Thus, fp2 might affect the deposition and patterning of microfibrils, the biosynthesis and deposition of cell wall components, which influences the formation of primary and secondary cell walls, the thickness of cell wall, cell elongation and expansion, plant morphology and plant strength in rice.

The high molecular weight glutenin subunit (HMW-GS) pair 1Bx13+1By16 are recognized to positively correlate with bread-making quality; however, their molecular data remain unknown. In order to reveal the mechanism by which 1By16 and 1Bx13 creates high quality, their open reading frames (ORFs) were amplified from common wheat Atlas66 and Jimai 20 using primers that were designed based on published sequences of HMW glutenin genes. The ORF of 1By16 was 2 220 bp, deduced into 738 amino acid residues with seven cysteines including 59 hexapeptides and 22 nanopeptides motifs. The ORF of 1Bx13 was 2 385 bp, deduced into 795 amino acid residues with four cysteines including 68 hexapeptides, 25 nanopeptides and six tripeptides motifs. We found that 1By16 was the largest y-type HMW glutenin gene described to date in common wheat. The 1By16 had 36 amino acid residues inserted in the central repetitive domain compared with 1By15. Expression in bacteria and western-blot tests confirmed that the sequence cloned was the ORF of HMW-GS 1By16, and that 1Bx13 was one of the largest 1Bx genes that have been described so far in common wheat, exhibiting a hexapeptide (PGQGQQ) insertion in the end of central repetitive domain compared with 1Bx7. A phylogenetic tree based on the deduced full-length amino acid sequence alignment of the published HMW-GS genes showed that the 1By16 was clustered with Glu-1B-2, and that the 1Bx13 was clustered with Glu-1B-1
alleles.

In vitro Transient Expression System of Latex C-serum was used for Analysis of Hevein Promoter in Response to Abscisic Acid in Hevea brasiliensisAuthor: Xiao-Wen Fei and Xiao-Dong Deng
Journal of Integrative Plant Biology 2008 50(3): 338-344DOI: 10.1111/j.1744-7909.2007.00628.x

Hevein has been found to be an essential element in coagulation of rubber particles in latex of rubber trees. In a previous study, we cloned a 1 241-bp fragment of a 5' upstream region of the hevein gene by genome walking. This fragment was analyzed by a 5' end nested deletion method in the present study, fused with a uidA (gus) gene to produce a series of tested constructs, which were transferred into C-serum of latex and the Gus activities were detected. Results showed that the fragment from −749 to −292 was sufficient for expression of gus gene in latex, and the fragment from −292 to −168 was crucial in response to abscisic acid inducement. In a transient transgenic test of rubber leaf with particle bombardment, construct Hev749 conferred gus-specific expression in veins, in which the latex tubes mainly distributed. This implies that the fragment from −749 to −292 was laticiferous-specific.

To identify alien chromosome in recipient progenies and analyze genome component in polyploidy, genomic in situ hybridization (GISH) technique suitable for cotton was developed using increased stringency conditions in the study. The increased stringency conditions were a combination of the four factors in the following optimized state: 100:1 ratio of blocking DNA to probe, 60% formamide wash solution, 43∼C wash temperature and a 13 min wash. Under these specific conditions using gDNA from Gossypium sturtianum (C1C1) as probe, strong hybridization signals were only on chromosomes from C1 genome in somatic cells of the hybrid F1 (G. hirsutum G. sturtianum) (AtDtC1). So, GISH was able to discriminate parental chromosomes in the hybrid. Further, we developed a multi-color GISH to simultaneously discriminate the three genomes of the above hybrid. The results repeatedly displayed the three genomes, At, Dt, and C1, and each set of chromosomes with a unique color, making them easy to identify. The power of the multi-color GISH was proven by analysis of the hexaploid hybrid F1 (G. hirsutum G. australe(AtAtDtDtG2G2). We believe the powerful multi-color GISH technique could be applied extensively to analyze genome component in polyploidy and identify alien chromosomes in the recipient progenies.

The variation of 90 Laminaria gametophyte clones representing the introduced Laminaria japonica (Group 1) and Laminaria longissima (Group 2), the varieties of L. japonica (Group 3) and the varieties derived from interspecific hybrids (Group 4) was determined with 18 microsatellite markers. The allelic diversity and Nei's gene diversity of Group 1 were significantly higher than those of Group 2 (2.9 vs. 1.8 and 0.414 vs. 0.161, respectively), demonstrating that the variation of the introduced L. japonica is richer than that of L. longissima. Both allelic diversity and Nei's gene diversity of Group 3 were lower than those of Group 1, indicating that only a portion of variation of L. japonica was incorporated into the varieties of L. japonica. Significant genetic differentiation was detected between four groups and between female (Population 1) and male (Population 2) gametophyte clones in each group. The variation among groups accounted for 39.95%, while that among populations accounted for 21.65% of the total. The genetic distance between Group 1 and Group 4 was obviously longer than that between Group 2 and Group 4 (0.686 vs. 0.291), indicating that maternal gametophyte clone contributed more variation to the hybrids than the paternal gametophyte clone did.

The complete process of meiosis was investigated in Pinus wallichiana Jacks, Pinus strobus L. and their artificial hybrid (F1) using microsporocytes. It is revealed that there were slightly lower chiasma frequency, lower ring bivalent frequency, lower meiotic index and distinctly higher frequency of aberrance (chromosomal bridges, fragments or micronuclei) in pollen mother cells (PMCs) of the hybrid (F1) than those of the parental species, which show a certain degree of differentiation between homologous chromosomes of the two parents. However, relatively higher frequency of ring bivalents and higher meiotic index in all the three entities indicate the great stability of genomes of parental species, and the differentiation of genomes between the two parents must have been slight. Total nineteen signal loci of 18S rDNA were observed in nine bivalents of the hybrid (F1), among which one bivalent bears two loci, while the others have only one. It is suggested that distinct differentiation at genetic level existed in homologous chromosomes of the two parental species, whereas only slight differentiation at karyotypic and genomic levels take place between the parent species.

Variations in the trnK region of chloroplast DNA were investigated in the present study using polymerase chain reaction每restriction fragment length polymorphism to detect the genetic structure and to infer the possible glacial refugia of Ginkgo biloba L. in China. In total, 220 individuals from 12 populations in China and three populations outside China were analyzed, representing the largest number of populations studied by molecular markers to date. Nineteen haplotypes were produced and haplotype A was found in all populations. Populations in south-western China, including WC, JF, PX, and SP, contained 14 of the 19 haplotypes and their genetic diversity ranged from 0.771 4 to 0.867 6. The TM population from China also showed a high genetic diversity (H = 0.848 5). Most of the genetic variation existed within populations and the differentiation among populations was low (GST = 0.2). According to haplotype distribution and the historical record, we suggest that populations of G. biloba have been subjected to extensive human impact, which has compounded our attempt to infer glacial refugia for Ginkgo. Nevertheless, the present results suggest that the center of genetic diversity of Ginkgo is mainly in south-western China and in situ conservation is needed to protect and preserve the genetic resources.

Single pollen grain polymerase chain reaction (PCR) has succeeded in several species, however only limited numbers of pollen grains were involved due to difficulties in pollen isolation and lysis. This has limited its application in genetic analysis and mapping studies in plants. A high-throughput (HT) procedure for collecting and detecting genetic variation in a large number of individual pollen grains by PCR is reported. The HT procedure involved the collection of individual pollen grains by a pair of special forceps and the lysis of pollen grains in a heated alkali/detergent solution followed by neutralization with a tris-ethylenediamine tetraacetic acid (TE) buffer. These resulting template solutions yielded PCR reactions involving the 5S ribosomal RNA intergenic spacers, randomly amplified polymorphic DNA, and simple sequence repeats markers. Using this procedure, one person with experience could collect and process up to 288 single pollen grain PCR reactions per day. The method worked well on sugarcane, corn, Miscanthus spp., snap bean, sorghum, and tomato. The ability to collect and conduct PCR on individual pollen grains on a large scale offers a new approach to genetic analyses and mapping studies in an easily controllable environment with a considerable cost reduction. The method will also significantly benefit studies in species that are difficult subjects for classical genetic research.